ES2397178B2 - PROCEDURE FOR OBTAINING BIOFERTILIZERS AND BIO STIMULANTS FOR AGRICULTURE AND ANIMAL FEEDING. - Google Patents

Abstract

Method for obtaining biofertilizers and biostimulants for agriculture and animal feed. # The invention defines a method for obtaining an organic enzyme extract from residues from the manufacture of beer that is made in a single container and comprising the steps of: ( a) add a concentrated base to the residues in suspension form to adjust their pH; (b) subjecting the mixture obtained in (a) to a pressure higher than atmospheric pressure and to an elevated temperature; and (c) subjecting the mixture obtained in (b) to an enzymatic hydrolysis to obtain an organic enzyme extract. This procedure allows to obtain extracts with a high biofertilizing and biostimulant capacity and a high capacity for bioabsorption by animals and plants, so they are especially useful in organic farming and animal feed.

Description

PROCEDURE FOR OBTAINING BIOFERTILIZERS and BiosTIMULANTS FOR AGRICULTURE AND ANIMAL FEEDING

FIELD OF THE INVENTION

The invention relates to the field of chemistry applied to agriculture, more specifically to organic plant nutrition, and animal feed. In particular, the invention relates to a process for obtaining organic enzyme extracts from residues of the beer industry that can be used as biostimulants and biofertilizers in agriculture, particularly organic farming, and as protein supplements in animal feed.

BACKGROUND OF THE INVENTION

As is well known in the state of the art, a fertilizer is an organic or inorganic substance, natural or synthetic, which is used to enrich the soil and provide plants with one or more of the nutritional elements essential for their normal vegetative development.

In addition to being a complete source of nutrients for plants, organic fertilizers provide organic matter to the soil that, for sandy soils or with low clays

activity,

by example, It represents a improvement in the

properties

physical, chemical Y biological, by its effect

conditioner .

Sayings fertilizers they can contain

Micronutrients and beneficial microorganisms such as yeasts, bacteria or fungi.

Thus, for example, Kobayashi et al. have studied the effect of a Saccharomyces cerevisiae yeast extract combined with an inorganic fertilizer on the growth of pea plants ("Effect of yeast extracts on higher plants" Plant

and Soil (1980) f 57 (1) f 41-7; "Effect of addition of the materials autolyzed from yeast cells on plant growth" Kankyo Kagaku Sogo Kenkyusho Nenpo (1980), 7, 87-90).

Similarly, Kudryasheva et al. ("Amino acids and their utilization, Pishchevaya Promyshlennost (Moscow, Russian Federation) (1992) I (4) I 13-14) have conducted a study on the use of amino acids from different strains of yeast, particularly bread yeast Saccharomyces cerevisiae, in fertilizers, food additives, etc.

In document HU 9902060 an aqueous fertilizer composition for the leaves and roots of plants containing Saccharomyces yeast is described, in addition to trace elements, complexing agents, buffering agents and other nutrient series such as amino acids, humic acids, enzymes, sugars, etc

More recently, document CN 1966663 ("COMPOSITE MICROORGANISM FOLIAGE FERTILIZER BACTERIA AGENT ANO ITS PRODUCING METHOD AND USE", 05/23/2007) describes a microbial agent that is composed of Saccharomyces cerevisiae, Lactobacillus plantarum, Mucor racemosus and Aspergillus or Aspergillus or Aspergillus or Aspergillus or Aspergillus or Aspergillus or Aspergillus , and which is used as a foliar fertilizer to promote the growth of root and leaf plants by improving cultivated soils and increasing crop yields.

Likewise, CN 19191800 describes a nutrient for animals and plants that is prepared by diluting Saccharomyces cerevisiae yeast sludge in water until an emulsion is obtained; mixing this with papain, neutral proteinase and sodium chloride; hydrolyzing the mixture afterwards; inactivating enzymes; and finally, concentrating or drying the product

obtained. The final product contains a large amount of nutrients high absorption speed.

Document 1919870 describes a plant nutrient that improves soil microorganisms, promotes plant growth, increases fertilizer utilization rates and increases resistance of plants to disease and stress. This nutrient comprises the yeast Saccharomyces cerevisiae, the bacteria Lactobacillus plantarum and Lactobacillus acidophilus, and other components such as potato or coffee derivatives, glucose, peptone, magnesium and manganese sulfates, dipotassium hydrogen phosphate and sodium chloride.

In US 2003022357 a biological fertilizer based on yeast cells of the genus Saccharomyces and sludges from storage plants is described

or wastewater treatment, in which the yeast cells are activated by the application of a magnetic field.

Likewise, document US 2002187900 describes a biological fertilizer comprising yeast cells of the genus Saccharomyces activated electromagnetically combined with purines from cattle. Similarly, US2002187552 discloses a biological fertilizer that also comprises electromagnetically activated Saccharomyces cells combined with bird slurry.

Through enzymatic technology fertilizers have been obtained using other organic waste such as plant waste from parks and gardens, animal waste, agroindustrial waste and solid waste

domiciliary

Agroindustrial activities produce a large amount, only in Spain of the order of millions of Tm / year, of agroindustrial organic by-products (SOA) resulting from activities as diverse as the wine industry, beer, meat production, flour, rice, dairy , etc . The generation of large quantities of by-products is one of the main problems created, due to their accumulation in the environment, and today there is great political and social pressure to reduce pollution.

In spite of their diverse origin, all these agro-industrial by-products are characterized by their high organic matter content, being especially very rich in proteins. This determines that the protein reservoirs with the greatest application potential in the food industry, in the pharmaceutical industry and also in the agrochemical (organic) industry, which uses them to obtain nutritional products for plants, as well as for soil (nitrogen source)

In order to improve the properties of proteins and thus achieve greater use, in recent years the protein extraction processes have been developed and optimized, such as hydrolysis processes, which improve their functional quality.

Thus, document US 5, 393, 318 A ("A METHOD OF PRODUCING ORGANIC FERTILIZER BY USING FISH AS RAW MATERIAL"; 28/0211995) refers to a method for producing organic fertilizer by using fish as raw material. The method comprises several stages: fish cooking and subsequent elimination of fish.

solids and fats to obtain a stock solution, which will be subjected to enzymatic hydrolysis, giving a clarified liquid that can be used as a fertilizer.

Document US 20020129631 Al ("METHOD FOR THE PREPARATION OF A PROTEIN HYDROLYZATE USABLE AS A FERTILIZER"; 07/03/2002) describes a method for achieving a

hydrolyzed

protein Useful how fertilizer to split from the

cooking

Y pressing from waste from Butcher shop applying two

enzymatic hydrolysis.

WO 2006/082264 ("METHOD OF OBTAINING BIOSTIMULANTS FROM AGRO-INDUSTRIAL RESIDUES"; 08/08/2006) refers to a method of obtaining plant biostimulants from agroindustrial residues such as wastes from the carob industry or those of the bioalcohol industry from corn seeds, for example. The method contemplates a first fermentative stage in which the secretory microorganisms of hydrolytic enzymes are produced, where the culture medium is the residue and a second phase in which these fermentation broths of the residues are used as hydrolytic instruments to treat more of the same waste.

Also, document US 2007/0039362 Al ("PROGRESSIVE DIGESTION PROCESS FOR PRODUCING FERTILIZER"; 02/22/2007) shows a process of progressive enzymatic digestion for the production of fertilizers from a mixture of products rich in organic matter such as manure, food waste from the food industry or landfill, vegetable processing remains or grass clippings. This progressive digestion consists of two first mesophilic episodes and a third thermophile, with separation of

solid.

Also in document US 2008/0302151 Al ("SOLUBLE LIQUID FERTILIZER FOR ORGANIC AGRICULTURE DERIVED FROM SOY MEAL"; 11/12/2008) a manufacturing process of an organic fertilizer by enzymatic hydrolysis of soybean meal, with elimination of the insolubles resulting from said hydrolysis.

Patent 1'10 2010/114203 ("METHOD FOR MANUFACTURING AMINO ACID LIQUID FERTILIZER USING LIVESTOCK BLOOD AND AMINO ACID LIQUID FERTILIZER MANUFACTURED THEREBY"; 07/10/20101 describes a method of producing a liquid amino acid fertilizer by enzymatic hydrolysis cattle .

On the other hand, Celus et al. (CELUS, l. Et al. "ENZYMATIC HYDROLYSIS OF BREWER 'S SPENT GRAIN PROTEINS AND TECHNOFUNCTIONAL PROPERTIE $ OF THE RESULTING HYOROLYSATES", J. Agric. Food Chem., Sept. 2007, Vol. 55 No. 21, pages 87038710) describe Enzymatic hydrolysis of depleted brewery or bagasse (BSG) grain proteins, which is the insoluble residue of barley malt resulting from the manufacture of the must prior to alcoholic fermentation of the same for brewing, and constituting The main byproduct of the beer industry. The resulting hydrolysates, because of their good foaming and emulsifying properties, are used in the food and beverage industry.

Likewise, document ES 2329750 Al ("PROCEDURE FOR OBTAINING A FERTILIZING PRODUCT FROM WASTE MANUFACTURING RESIDUES", HEINEKEN ESPAÑA SA, 11/30/2009) describes a process in several stages of manufacturing a fertilizer from residues from the manufacture of beer after the alcoholic fermentation of the must. Said process is a multi-stage process with phase separation in which the residues are alkalized, subjected to separation to obtain a liquid phase, which is subsequently subjected to an acid treatment and from which, in turn, a solid phase that finally undergoes enzymatic hydrolysis or acid hydrolysis. This procedure, however, has the disadvantages of being complex and expensive and, in addition, can produce acid waste. The fertilizer product obtained has a low yield and can contain a large amount of substances that cannot be absorbed by plants and potentially toxic.

There continues to exist in the state of the art, therefore, the need to take advantage of the residues of the beer industry obtained after most of the alcoholic fermentation or in a simpler and more profitable way.

Surprisingly, the present inventors have discovered that the physical treatment of all leaving residues without phase separation at a pressure higher than the atmospheric and at an elevated temperature, once treated with a concentrated base and before being subjected to them At enzymatic hydrolysis, it is possible to obtain, in a simpler, less expensive and less continuous way, enzymatic organic extracts containing practically all the hydrolyzed protin in the form of free amino acids, oligopeptides and other peptides of greater molecular weight and, moreover, practically All nutrients from the starting residue. Such extracts, therefore, have a greater biofertilizing and biostimulant capacity, as well as a greater capacity for bi-absorption by animals and plants. Therefore, these products are especially useful in

ecological agriculture

Y in feeding animal, how additives

nutritional

from tall value added for won or for

aquaculture,

for example .

OBJECT OF THE INVENTION

The present invention, therefore, is intended to provide a process for obtaining organic enzyme extracts from residues from beer manufacturing.

On the other hand, the object of the invention is to provide the organic enzymatic extracts obtainable by said process.

Finally, the object of the invention is to provide the use of said organic extracts in agriculture and animal feed.

DESCRIPTION OF THE FIGURES

Figure 1 shows the block diagram of a particular embodiment of the process of the invention, in which the residues of beer brewing in suspension form in beer:. :: a are subjected to alkaline LraLamienLo, followed by a treatment physical and enzymatic hydrolysis

to directly obtain an enzymatic organic extract (HEOC), as well as, after additional stages of concentration and / or separation, other enzymatic organic tracts of interest

(HEO-C concentrate, EOEO- $ and EOEO-S concentrate) and insoluble organic matter (MOl-O).

Figure 2 shows the block diagram of a particular embodiment of the process of the invention, in which the residues of the beer manufacturing in the form of

The suspension in beer is separated from it and then resuspended with water before being subjected to alkaline treatment, followed by physical treatment and enzymatic hydrolysis to directly obtain an organic enzyme extract (HEP5 e), as well as, after additional concentration steps and / or separation, other enzymatic organic extracts of interest

(HEP-C concentrate, EOEP- $ and EOEP-S concentrate) and subject matter

organic insoluble (MOI-P).

10 Figure 3 shows the peptide profile of one of the x tracts obtained by the process of the invention.

Figure 4 shows the production in grams of Cherry tomato as a function of time, when a control composition and two extracts obtained by the process of applying are applied

the invention .

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a method for

20 to obtain an organic enzyme extract from residues from beer manufacturing (hereinafter "the process of the invention"), which comprises the following steps:

(a) add a concentrated base to the residues in suspension form to adjust their pH;

(b) subjecting the mixture obtained in (a) to a pressure greater than 1 at atmospheric pressure and at an elevated temperature; Y

(e) subjecting the mixture obtained in (b) to an enzymatic hydrolysis to obtain an enzyme extract

organic and that is done in a single container.

In the context of the invention, the term "residues from the manufacture of beer" refers to the by-products obtained during the industrial brewing process. More particularly this waste

5 are made up of yeasts and by-products from the alcoholic fermentation of barley malt (proteins, water-soluble vitamins of the B complex, phosphates, minerals such as potassium, sulfur, magnesium, calcium or sodium, unsaturated fatty acids, lecithins, cephalins such carbohydrates

10 such as glycogen, trehalose, glucans or mannans, ethyl alcohol, carbon dioxide, traces of esters, aldehydes, ketones and higher alcohols, etc.). These wastes may be suspended in beer, as they leave the industrial plant, without the need for drying or concentration, or

15 be in solid form once the remaining beer has been removed. The remaining beer can be separated by any conventional method such as filtration, sedimentation or centrifugation, for example.

In a particular embodiment of the process of the invention, the elementary composition of the waste to be treated, expressed as a percentage by weight, is as follows:

% e

% N % P % K % S

Waste

suspended 44, 22 7, 73 1, 48 1, 78 0, 36

in

beer

Waste

without beer

remandant

47, 06 8, 56 1, 22 O, 65 0, 36

In the case of starting solid waste without beer

On the remaining side, a suitable solvent is added thereto in order to obtain a suspension that can be subjected to the process of the invention.

Thus, in a particular embodiment of the process of the invention, this comprises a stage prior to step (a) in which the residues are obtained in the form of a suspension. In a preferred embodiment, in this previous step the solvent is added to the solid waste until a suspension with a dry matter concentration of 8-25% by weight, preferably 15% by weight, is obtained.

In another preferred embodiment, the solvent used to suspend solid waste is water. In another preferred embodiment, the solvent used to suspend solid waste is a liquid solution or a suspension with organic matter from any agri-food industry that has a dry matter content of less than 10% by weight, preferably 4-8% by weight. . In a more preferred embodiment, the solvent used to suspend solid waste is whey from cheese making. In an even more preferred embodiment, the solvent used to suspend the solid residues is whey from the production of cheeses with a dry matter content of less than 10% by weight, preferably 4-8% by weight.

On the other hand, in the context of the invention the expression "in a single container" (one-pot) refers to the fact that the process is carried out without intermediate stages of separation so that the use of the starting waste is total, is that is, the nitrogen and mass yield is 100%.

Likewise, in the context of the invention the expression "biofertilizers and biostimulants" refers to compounds capable of stimulating the growth and development of plants and crops, as well as increasing and enhancing the

soil microbiological activity.

The residues in suspension form are subjected to the alkaline treatment of step (a) of the process of the invention, for which a suitable base selected by the expert is used. Thus, in a particular embodiment of the process of the invention, the base of step (a) is selected from ammonium hydroxide, potassium hydroxide and calcium hydroxide. In a preferred embodiment, the base is ammonium hydroxide. In another preferred embodiment, the base is potassium hydroxide. Said base is used in a concentrated form so as not to increase the reaction volume too much which would increase the energy costs of heating, concentrating, etc. , as well as equipment costs. Said concentration will be selected by the expert based on the pH range that he wishes to obtain in order to carry out the subsequent enzymatic hydrolysis. In a more preferred embodiment, 28% by weight ammonium hydroxide is used. In another more preferred embodiment 10 M potassium hydroxide is used.

After the alkaline treatment of the residues in the form of a suspension, the mixture is physically treated at a pressure higher than atmospheric pressure and at an elevated temperature.

Thus, in a particular embodiment of the process of the invention, in step (b) a pressure of 102-141 kPa is applied at an elevated temperature. In a preferred embodiment, in step (b) a pressure of 105-120 kPa is applied at an elevated temperature. In a more preferred embodiment, in step (b) a pressure of 115 kPa is applied at an elevated temperature.

Thus, in a particular embodiment of the process of the invention, in step (b) a pressure greater than atmospheric pressure is applied at a temperature of 90-140 oc. In a preferred embodiment, in step (b) a pressure is applied

5 higher than atmospheric pressure at a temperature of 100-130 oC. In a more preferred embodiment, in step (b) a pressure greater than atmospheric pressure is applied at a temperature of 120 oc.

In another particular embodiment of the process of the invention, in step (b) a pressure of 102-141 kPa is applied at a temperature of 90-140 oC. In a preferred embodiment, in step (b) a pressure of 105-120 kPa is applied at a temperature of 100-130 oC. In a more preferred embodiment, in

15 step (b) a pressure of 115 kPa is applied at a temperature of 120 oc.

This physical treatment will be carried out in any suitable device selected by the expert, such as an autoclave, for example.

After the physical treatment of the process of the invention, the enzymatic treatment is carried out. Optionally, after physical treatment and before enzymatic treatment

A concentrated base is added so that the mixture to be treated has the optimum pH value of the enzyme to be used.

The enzyme to be used in the enzymatic hydrolysis of the stage

(e) is a proteolytic enzyme of microbial, plant or

30 animal Thus, the enzyme to be used in the process of the invention can be an alkaline protease such as subtilisin, a neutral protease produced by Bacillus sp. , trypsin or chymotrypsin, for example.

Thus, in a particular embodiment of the process of the invention, the enzyme employed in step (e) is an alkaline protease. In a preferred embodiment, the enzyme employed in step (e) is subt ilisin.

In another particular embodiment of the process of the invention, the enzymatic hydrolysis of step (e) is carried out using a concentration of 0.05% -0.5% by volume of an enzyme stock solution with an activity of 70. 000 units

(azocasein assay). In a preferred embodiment, the enzymatic hydrolysis of step (e) is carried out using a concentration of 0.3% by volume of said stock solution.

On the other hand, the conditions of pressure, temperature, pH and time of enzymatic hydrolysis will be those in which the maximum activity of the enzyme is achieved. Thus, in another particular embodiment of the process of the invention, the enzymatic hydrolysis of step (e) is carried out at a temperature of 40-70 oC and at a pH of 8-11 for a time of 2-48 h. In a preferred embodiment, the enzymatic hydrolysis of step (e) is carried out at a temperature of 55 oC and at a pH of 9.2 for a period of 24 h.

In a particular embodiment of the process of the invention, during the enzymatic hydrolysis the pH value is kept constant by the addition of a base, such as, for example, ammonium hydroxide or potassium hydroxide.

The enzymatic treatment can be carried out in any suitable device selected by the expert, such as a reactor with temperature control and stirring, for example.

After this final enzymatic hydrolysis an organic enzyme extract is obtained, hereinafter "organic enzyme extract of the invention", which contains practically all the hydrolyzed starting protein, that is, more than 90% in

5 weight, in the form of free amino acids, oligopeptides and other peptides of greater molecular weight and, also, practically all the nutrients of the starting residue and, where appropriate, the nutrients of the liquid solution or suspension of organic matter of the agri-food industry employed in the

10 suspension of solid waste without remaining beer.

Thus, in another aspect of the invention an organic enzyme extract obtained by the process of the invention described above is provided. Said extract is

15 rich in proteins, mostly in the form of high bioabsorption free peptides and amino acids (free amino acids, oligopeptides and low molecular weight peptides with a size less than 10,000 daltons, as well as carbohydrates, and is fat and cholesterol: its chemical composition

20 nutritional in dry weight is as follows:

Proteins 45-55% Carbs 40-45% Ashes Rest

The amino acids, oligopeptides and peptides of low molecular weight that constitute it are nutritious substances that are easily absorbed and assimilated for plants, both foliar and root, being transported to plant organs such as buds, flowers or fruits, for example ( Gjalakshimi et al., 2004; Bioresource Technlogy (92): 291-296; Parrado et al., 2008 Bioresource Technology 99 (2008) 2312-2318). There can be 30 employees for the plant to make its own proteins,

saving a series of energy-consuming metabolic processes (Higgings, CF, Payne, JW, 1982. Plant peptides. In: Boulderf D. I Parthierf B. I (Eds). Encyclopedia of Plant Physiology 14A. Springer. 438-458) Y Not only can they be absorbed and used by plants, the existing microfauna in the soil environment is able to use them as a nitrogen source, essential for their growth and activity. This nitrogen is a fundamental element, since it is a basic constituent of proteins, nucleic acids, chlorophylls, etc. and, therefore, allows the development of the metabolic activity of plants and microorganisms. Likewise, amino acids, oligopeptides and low molecular weight peptides are also easily absorbable and assimilable by animals when they are incorporated into their diet.

Therefore, the organic enzyme extract of the invention can be used as such in agricultural and livestock applications, for example.

Thus, in another aspect of the invention, the use of the organic enzyme extract previously described in agriculture and animal feed is provided. More particularly, the organic enzyme tract of the invention can be used as a biostimulant and biofertilizer in organic farming given its special composition of free amino acids, oligopeptides and low molecular weight peptides. It can also be used as a nutritional additive of high added value for animal feed, more particularly, in feed for

animals

in cattle raising (bovine, sheep goat etc. ) or

aquaculture,

or for animals domestic or of company for

example .

The method of applying the organic enzyme extract of the invention to an agricultural crop can be carried out by

any

technique conventional such how, by example,

application

direct in he ground, or by via foliar or by

fertirrigation

On the other hand, the organic enzyme extract of the invention may alternatively be subjected to subsequent stages of concentration and / or separation for stabilization.

In a particular embodiment, the process of the invention comprises a stage after step (e) in which the organic enzyme extract obtained after said step

(e) is subjected to concentration to obtain a concentrated organic enzyme extract, hereinafter "concentrated organic enzyme extract of the invention". The concentrated organic enzyme extract of the invention has at least 40% by weight dry matter, preferably at least 50% by weight dry matter and, more preferably, 50-55% by weight dry matter. This concentration can be carried out by any conventional method of the prior art, such as, for example, by heating and using a rotary evaporator with thermostated bath or a reverse osmosis system, for example, or another suitable device.

In another particular embodiment, the process of the invention comprises a stage after step (c) in which the organic enzyme extract obtained after said step

(c) is subjected to separation to obtain: (i) a soluble organic enzyme extract, hereinafter "soluble organic enzyme extract of the invention", and (ii) a solid phase, hereinafter "insoluble organic matter of the invention". This separation can be carried out by any conventional method of the prior art, such as, for example,

.

by filtration or centrifugation using a decanter or other suitable industrial device, for example.

The soluble organic enzyme extract of the invention contains virtually all of the hydrolyzed starting protein, that is, more than 90% by weight, and therefore has a composition that also makes it suitable for use in agricultural and livestock applications. More particularly, it can be used as a biostimulant and biofertilizer in organic farming given its special composition of free amino acids, oligopeptides and low molecular weight peptides. It can also be used as a nutritional additive of high added value for animal feed, more particularly, in animal feed in livestock (bovine, sheep, goats, etc.) or aquaculture, or for domestic or companion animals, for example.

The insoluble organic matter of the invention, in turn, can be subjected to a final drying process to obtain a solid by-product in the form of a paste with a moisture content of 10-15% by weight. Said drying can be carried out by any conventional method, such as the use of circulating air drying ovens, for example.

The insoluble organic matter of the invention, concentrated

or without concentrating, it can also be used as a nutritional additive of high added value for animal feed, more particularly, in animal feed in livestock (bovine, sheep, goats, etc.) or aquaculture, or for domestic animals

or company, for example.

Optionally, the soluble organic enzyme extract of the invention can subsequently be subjected to concentration.

Thus, in a preferred embodiment, the process of the invention comprises a stage after step (e) in which the organic enzyme extract obtained after said step

(e) undergoes separation followed by a stage of

5 concentration of the soluble organic enzyme extract obtained to obtain a concentrated soluble organic enzyme extract, hereinafter "concentrated soluble organic enzyme extract of the invention", The concentrated soluble organic enzyme extract of the invention has at least 40% in

10 weight of dry matter, preferably at least 50% by weight of dry matter and, more preferably, 50-55% by weight of dry matter. This concentration can be carried out by any conventional method of the prior art, such as, for example, by heating and vacuum using a

15 rotary evaporator with thermostated bath or a reverse osmosis system, for example, or another suitable device.

The concentrated organic enzyme extract of the invention and the concentrated soluble organic enzyme extract of the invention are extracts with a composition that also makes them suitable for use in agricultural and livestock applications. More particularly, they can be used as biostimulants and biofertilizers in organic farming

given its special composition of free amino acids,

25 oligopeptides and low molecular weight peptides. They can also be used as a nutritional additive of high added value for animal feed, more particularly, in animal feed for livestock (bovine, sheep, goats, etc.) or aquaculture, or for domestic or companion animals, by

30 example.

The following examples illustrate the invention and should not be considered as limiting its scope.

EXAMPLE 1

Obtaining extracts from a beer manufacturing residue in the form of a suspension in beer.

The block diagram of this particular embodiment of the process of the invention is shown in Figure 1.

500 9 of a beer residue in the form of a beer suspension (RFC) was treated in an open glass reactor with 28% NH3 to a pH of 9.6 with stirring (60-80 rpm). The dry matter percentage of the RFC was 10, 27% w / w (10, 77% w / v) and its chemical composition was as follows:

Table 1 Elemental chemical composition of the RFC (% by weight)

% e

% N % P % K % S

44, 22

7, 73 1, 48 1, 78 0, 36

After the alkaline treatment, a physical process was carried out at a pressure of 115 kPa and at a temperature of 120 oc for 20 minutes in an autoclave. The solution thus obtained was adjusted to a pH of 9.2 with 10 M potash.

This solution at pH 9.2 was maintained at 55 ° C in a thermostated bath with stirring (60-80 rpm) and 0.3% v / v Subtilisin (stock protease solution 70,000 Activity Units was added / Azocasein test). The pH was maintained with 28% ammonia. These hydrolysis conditions were maintained for 24 hours.

The resulting liquid organic enzyme extract (HED-C) was collected at 24 hours, its composition being as follows:

Table 2 Elemental chemical composition of HED-C (% by weight)

% e

% N % P % K % S

42, 73

7, 1 1, 27 3, 88 0, 34

Table 3 Nutritional chemical composition in dry weight

Protein

47%

Carbohydrates

46%

Ashes

6%

5 The nitrogen and mass yield of this process is 100%, since there is no phase separation and the use of RFC is total.

Then 500 ml of HED-C were concentrated 5 times in

10 a rotary evaporator with a thermostated bath at 70 ° C, obtaining a concentrated organic enzyme extract (concentrated HED-C) in the form of a stable syrup with 50-55% by weight of dry matter.

On the other hand, 500 ml of HED-C was centrifuged at 4000 G

15 for 30 minutes. 400 mI of a soluble organic enzyme extract (EOEO-S, 80% of the initial volume) and 107.5 g of insoluble organic matter (MOl-O) were obtained, with dry matter content by weight of 9, 77 and 20 , 35%, respectively. The compositions are shown in Table 4 below.

20 chemicals of both products in dry matter. The mass yield of obtaining the EOEO-S is 65%, while the MOI-D maintains 35% of the initial matter.

Table 4 Elemental chemical composition of EOED- $ and MOl-D 25 (% by weight) EOED-S

% e

% N % P % K % S

41, 86

8, 11 1, 32 4, 82 0, 37

MOI-D

% e

% N % p % K % S

44, 39

5, 16 1, 18 2, 08 0, 29

Table 5. EOEDS dry weight nutritional chemical composition

Protein

54%

Carbohydrates

39%

Ashes

6%

Likewise, the amino acid composition of EODE-S was determined, as well as its peptide profile. The results are shown in Tables 6 and 7.

10 Table 6 Amino acid composition of EOED- $ (% by weight)

•from

•from

amino acids

amino acids

Isoleucine

2, B9 To the girl 5, 9

Leucine

4, 87 Arginine 1, 9

Lysine

8, 81 Asparagine 4, 85

Cysteine

+ Methionine 0, 67 Aspartic 2, 78

Phenylalanine

+ Tyrosine 3, 82 Glutamic 7, B

Threonine

2, 21 Glycine 3, 27

Tryptophan

0, 9B Hist idina 7, 29

Valine

5, 87 Serina 3, 04

Table 7 EOED- $ peptide profile (% by weight)

Size (Oalto ns)

% of the total

> 10. 000

38, 31

10. 000 5. 000

18, 07

3 . 000 1,000

12, 23

one . 000 300

6, 1 8

<300

25, 22

A graph with the peptide profile of the EOED-S extract is shown in Figure 3, in which peptides with a size less than 300 Da correspond to free amino acids and small peptides.

The concentration of EOED-S was then carried out, leaving 50.5 ml of a soluble organic enzyme extract

concentrated

(EOED-S concentrated) in shape from syrup with a

77 f

4% in weight from matter organic The MOI -D be dry to 90 oc

until

contain 25 9 with a 10-15% in weight of humidity .

EXAMPLE 2

Obtaining extracts from a beer manufacturing residue in the form of a suspension in water.

The block diagram of this particular embodiment of the process of the invention is shown in Figure 2, where it is based on a residue in the form of a suspension in water which, in turn, has been obtained from the beer residue in the form of a suspension in beer from which the remaining beer has been removed and to which water has subsequently been added.

Thus, 500 ml of a beer residue in the form of a beer suspension (RFC) was treated in a centrifuge at 4000 G for 1-2 hours keeping the temperature at 4 ° C.

The 354 ml of supernatant obtained (beer) I was extracted, leaving 149 g of a solid waste beer residue (RFC-Sol), with a dry matter percentage of 24, 30% by weight and whose composition is shown in Table 8 :

Table 8 Elemental chemical composition of RFC-sun (% by weight)

% C

% N % P % K % S

47, 06

8, 56 1, 22 0, 65 0, 36

At 149 9 of the RFC-Sol, 354 ml of water were added to the network, and it was homogenized, until the initial volume 5 (500 ml) was completed. It was then alkalized with ammonia until pH

9, 6 with agitation.

This solution was subjected to a physical process at a pressure of 115 kPa and at a temperature of 120 oc for 20 minutes in an autoclave. The solution thus obtained was adjusted to a pH of 9.2.

with 10 M potash.

This solution at pH 9, 2 was introduced into an open reactor and maintained at 55 ° C in a thermostated bath with stirring.

15 (60-80 rpm) and 0.3% v / v of Subtilisin (70,000 protease stock solution / Azocasein Assay Units) was added. The pH was maintained with 28% ammonia. These hydrolysis conditions were maintained for 24 hours.

The resulting organic enzyme extract (HEP-C) was collected, the composition being the following:

Table 9 Elemental chemical composition of HEP-C (% by weight)

% C

% N % P % K % S

45, 17

7, 80 1, 04 3. 4. 5 0, 34

25 Table 10 Dry chemical nutritional composition in dry weight

Protein

52%

Carbohydrates

41%

Ashes

6%

The nitrogen and mass yield of this process is 100%, since there is no phase separation and the use of RFC-Sol is total.

5 Next, 500 ml of HEP-C was concentrated in a rotary evaporator with a thermostated bath at 55 oC, obtaining a concentrated organic enzyme extract (concentrated HEP-C) in the form of a stable syrup with 50-55% by weight of dry matter .

On the other hand, 500 ml of HEP-C was centrifuged at 4000 G for 30 minutes.

In this way, 400 ml were obtained in the form of the soluble organic enzyme extract (EOEP-Sl, with 5.86% by weight of 15 dry matter (23.44 g) and 107.5 9 of insoluble organic matter

(MOI-P), with 19, 58% by weight dry matter (21 g).

The mass yield was 52, 81% in the EOEP-S and 47, 19% in the MOl-P. The nitrogen yield was 55% in 20 EOEP-S and 45% in MOl-P.

The chemical compositions of both products obtained are shown in Table 11.

25 Table 11. Elemental chemical composition of EOEP-S and MOI-P (% by weight) EOEP-S

% e

% N % P % K % S

42, 8

8, 784 1, 38 4, 34 0, 38

MOI-P

% e

% N % P % K % S

43, 83

6, 09 fifteen 1, 78 0, 32

The concentration was then carried out up to 10 times by volume of the EOEP-S (containing approximately 60% by weight of dry matter). The MOI-P was dried at 90 ° C to contain 10-15% by weight of moisture, obtaining 16, 75 9 thereof.

EXAMPLE 3

Application of the extracts obtained in example 1 as biostimulants and biofertilizers in the production of cherry tomatoes in the greenhouse.

Two of the extracts obtained in Example 1 (HED-C and EOED-S) were tested in the production of Cherry tomatoes in the greenhouse (controlled humidity and temperature conditions). The control was treated exclusively with water, using blond peat as support.

The groups tested were defined by trays with 5 pots each, and a different product was applied to each tray. The treatments began after the germination of the plants and their transplantation to the pot.

The doses used were 1.3 grams of HED-C / liter of irrigation water and 0.939 grams of EOED-S / liter of irrigation water, depending on the nitrogen content.

Plant height

Height growth in centimeters of the different groups tested was measured. The results are shown in Table 12:

Table 12 Average height of plants (cm)

Time (days)

Control HED-C EOED-S

one

20, 4 19, 4 19, 4

8

27, 4 27 I 6 31

twenty-one

3. 4 3. 4. 5 35, 3

35

40, 7 47 47, 7

5

All control group groups tested in height. surpassed statistically to the

Tomato production Table 13 shows the Cherry tomatoes produced per plant.

data from the grams from

10

Table 13 Weight of tomatoes Cherrv produced (q)

CTRL HED-C EOED-S

90 days O O O 105 days 44, 52 137, 50 146, 32 120 days 179, 78 272, 20 285, 88

These results are shown graphically in Figure 4, from which it is concluded that the application of the extracts of the invention induces a notable increase in the production of tomatoes.

2.

Claims (15)

l. Procedure to obtain an organic enzyme extract from manufacturing residues 5 of the beer, characterized in that it comprises the following steps: {by adding a base to the waste in suspension form concentrated to adjust their pH; (b) subject the mixture obtained in (a) to a pressure 10 higher than atmospheric pressure and at a temperature of 90-140 ° C¡ and (e) subjecting the mixture obtained in (b) to an enzymatic hydrolysis to obtain an organic enzyme extract; 15 and because it is done in a single container. 2. Method according to claim 1, characterized in that it comprises a stage prior to step (a) of obtaining the waste in the form of a suspension. 3. Method according to claim 1, characterized in that it comprises a post-stage stage (wherein the organic enzyme extract obtained is subjected to concentration to obtain an organic enzyme extract 25 concentrated with at least 40% by weight dry matter. 4. A method according to claim 1, characterized in that it comprises a stage after step (c) in which the organic enzyme extract obtained is subjected to 30 separation to obtain: (i) a soluble organic enzyme extract, and (iil a solid phase. 5. Procedure according to claim 4, characterized in that the soluble organic enzyme extract is subjected to concentration to obtain a soluble organic extract or enzyme concentrated with at least 40% by weight dry matter. Method according to claim 1, characterized in that the concentrated base of the stage (al is selected from ammonium hydroxide, potassium hydroxide and calcium hydroxide.

7. Process according to claim 6, characterized in that the concentrated base of the step (al is ammonium hydroxide.

8. Method according to claim 1, characterized in that in step (b) a pressure of 102-141 kPa is applied at a temperature of 90-140 ° C.

9. Method according to claim 8, characterized in that a pressure of 115 kPa is applied at a temperature of 120 oC.

10. Method according to claim 1, characterized in that the enzyme used in the e t apa (e) is an alkaline protease.

eleven . Method according to claim 10, characterized in that the enzyme used in step (e) is subtilisin.

12.

Method according to claim 1, characterized in that in step (e) the enzymatic hydrolysis is carried out at a temperature of 40-70 oC and at a pH of 8-11 for a time of 2-48 h.

13.

Method according to claim 12, characterized

because the enzymatic hydrolysis is carried out at a temperature of 55 oC and at a pH of 9.2 for a period of 24 h 14. Organic enzyme extract obtainable by the process according to any one of claims 1-13, characterized in that it contains all the protein of the hydrolyzed starting residues in the form of amino acids 10 free, oligopeptides and other small peptides of greater molecular weight. 15. Use of the organic enzyme extract according to claim 14 in agriculture and animal feed. fifteen